Influence of lead on the activity of soil microorganisms in two Beskidy landscape parks

The aim of the study was to assess the potential impact of lead on soil metabolism in two landscape parks localized in the Beskid Śląski and Beskid Żywiecki mountains which were affected, among others, by air pollution from the Upper Silesian Industrial Region, the largest industrial zone in Poland. The study was carried out in six locations with different lead levels in the soil environment. Each plot was equipped with four pairs of vacuum ceramic lysimeters to assess the mobility of Pb in the soil. The metabolic activity was assessed by measuring: soil enzyme activity, soil respiration and by studying community-level physiological profiling (CLPP) using Biolog EcoPlates technique. The soil to the examination was collected near the stands with the lysimeters from two soil horizons (A and B layer). The analyses carried out showed that the factors that had the greatest influence on lead mobility were the organic carbon content and the soil pH. The elevated lead level in the topsoil (layer A) could affect the functional biodiversity of soil microorganisms, but low soil pH was a more likely limiting factor. In the subsoil (layer B), lower lead content was found and its probable effect on soil microbial activity was small. In summary, it can be concluded that the assessment of the influence of heavy metals on soil metabolism is not easy, and the Biolog system has proven to be a sensitive tool for assessing the potential impact of heavy metals on the soil environment.

A High-Throughput Metabolic Microarray Assay Reveals Antibacterial Effects of Black and Red Raspberries and Blackberries against Helicobacter pylori Infection

Helicobacter pylori infection is commonly treated with a combination of antibiotics and proton pump inhibitors. However, since H. pylori is becoming increasingly resistant to standard antibiotic regimens, novel treatment strategies are needed. Previous studies have demonstrated that black and red berries may have antibacterial properties. Therefore, we analyzed the antibacterial effects of black and red raspberries and blackberries on H. pylori. Freeze-dried powders and organic extracts from black and red raspberries and blackberries were prepared, and high-performance liquid chromatography was used to measure the concentrations of anthocyanins, which are considered the major active ingredients. To monitor antibiotic effects of the berry preparations on H. pylori, a high-throughput metabolic growth assay based on the Biolog system was developed and validated with the antibiotic metronidazole. Biocompatibility was analyzed using human gastric organoids. All berry preparations tested had significant bactericidal effects in vitro, with MIC90 values ranging from 0.49 to 4.17%. Antimicrobial activity was higher for extracts than powders and appeared to be independent of the anthocyanin concentration. Importantly, human gastric epithelial cell viability was not negatively impacted by black raspberry extract applied at the concentration required for complete bacterial growth inhibition. Our data suggest that black and red raspberry and blackberry extracts may have potential applications in the treatment and prevention of H. pylori infection but differ widely in their MICs. Moreover, we demonstrate that the Biolog metabolic assay is suitable for high-throughput antimicrobial susceptibility screening of H. pylori.

Aspergillus niger Environmental Isolates and Their Specific Diversity Through Metabolite Profiling

We present a biological profile of 16 Aspergillus niger environmental isolates from different types of soils and solid substrates across a pH range, from an ultra-acidic (<3.5) to a very strongly alkaline (>9.0) environment. The soils and solid substrates also differ in varying degrees of anthropic pollution, which in most cases is caused by several centuries of mining activity at old mining sites, sludge beds, ore deposits, stream sediments, and coal dust. The values of toxic elements (As, Sb, Zn, Cu, Pb) very often exceed the limit values. The isolates possess different macro- and micromorphological features. All the identifications of Aspergillus niger isolates were confirmed by molecular PCR analysis and their similarity was expressed by RAMP analysis. The biochemical profile of isolates based on FF-MicroPlate tests from the Biolog system showed identical biochemical reactions in 50 tests, while in 46 tests the utilisation reactions differed. The highest similarity of strains isolated from substrates with the same pH, as well as the most suitable biochemical tests for analysis of the phenotypic similarity of isolated strains, were confirmed when evaluating the biochemical profile using multicriterial analysis in the Canoco program. The isolates were screened for mycotoxin production by thin-layer chromatography (TLC), as well. Two of them were able to synthesise ochratoxin A, while none produced fumonisins under experimental conditions. Presence of toxic compounds in contaminated sites may affect environmental microscopic fungi and cause the genome alteration, which may result in changes of their physiology, including the production of different (secondary) metabolites, such as mycotoxins.

Identification and Characterization of Metabolic Potential of Different Strains from Genus Rhizobium

Bacteria of the Rhizobium genus form a group of microorganisms existing in the environment in two forms: symbiotic, in the root nodules of Fabaceae sp. plants and free-living, and saprophytic in the soil environment. The subject of this study was genetic identification and characterization of metabolic activity of different strains from Rhizobium genus bacteria. The study was conducted on the 16 bacteria strains from the collection of the Department of Agricultural Microbiology, Institute of Soil Science and Plant Cultivation in Puławy, Poland. Based on the sequencing of PCR products, we found that all strains belong to one species—Rhizobium leguminosarum. The study of metabolic activity was performed using the GEN III BIOLOG system method (Biolog Inc., Hayward, CA, USA). Metabolism analysis of all R. leguminosarum strains with the use of GEN III™ plates showed that carbohydrates (CH) were the most intensively utilised group of substrates. Between the Rhizobium leguminosarum strains, there are metabolic differences in terms of the studied features.

Identification and Biochemical Characterization of some Strains of Clavibacter michiganensis subsp. Michiganensis, in Romania

Clavibacter michiganensis subsp. michiganensis is responsible for the ʻbacterial canker of tomato’. The study aimed to identify and characterize biochemically some strains of the bacteria. Five strains were used for biochemical identification and characterization and two reference strains were used as positive controls. The biochemical characterization was performed with Biolog Microbial Identification System. All the strains used many carbon sources and were positive for growth in the presence of: 1%NaCl, Nalidixic Acid, Aztreonam, Potassium Tellurite and pH 6, but negative for growth and sensitivite to: Troleandomycin, Lincomycin, Vancomycin, Fusidic Acid, Eimfamycin SV, Guanidine HCl, D-Serine, Minocycline and Niaproof 4. The differences between the Biolog system database and the studied strains did not change their classification at the level of genus and species, but modifed or did not allow the classification at the subspecies level. Furthermore, the analyzed strains had metabolic diversity.

The Dose of Fungal Aerosol Inhaled by Workers in a Waste-Sorting Plant in Poland: A Case Study

Bioaerosol monitoring is a rapidly emerging area in the context of work environments because microbial pollution is a key element of indoor air pollution and plays an important role in certain infectious diseases and allergies. However, as yet, relatively little is known about inhaled doses of microorganisms in workplaces. Today, the important issue of social concern is due to waste management, transport, sorting, and processing of wastes and their environmental impact and effects on public health. In fact, waste management activities can have numerous adverse effects on human wellbeing. Health effects are generally linked to exposure (EX), defined as the concentration of a contaminant and the length of time a person is exposed to this concentration. Dose is an effective tool for evaluating the quantity of a contaminant that actually crosses the body’s boundaries and influences the goal tissue. This document presents an analysis of the fungal waste-sorting plant EX dose (FWSPED) inhaled by workers in a waste-sorting plant (WSP) in Poland in March 2019. The main purpose of this research was to assess FWSPED inhaled by workers in two cabins at the WSP: the preliminary manual sorting cabin (PSP) and the purification manual sorting cabin (quality control; QSP). It was found that the FWSPED inhaled by workers was 193 CFU/kg in the PSP and 185 CFU/kg in the QSP. Fungal particles were quantitatively evaluated and qualitatively identified by the GEN III Biolog system. During the research, it was found that isolates belonging to the Aspergilus flavus and Penicillum chrysogenum strains were detected most frequently in the WSP. The total elimination of many anthropogenic sources is not possible, but the important findings of this research can be used to develop realistic management policies and methods to improve the biological air quality of WSPs for effective protection of WSP workers.

Carbon source utilisation and evaluation of the Biolog system in the identification of Actinobacillus pleuropneumoniae

Sixty-eight Actinobacillus pleuropneumoniae strains were isolated from porcine acute pleuropneumonia cases from different parts of Hungary between 2000 and 2014. A total of 41 isolates were identified as A. pleuropneumoniae bio-type I and 27 strains as biotype II based on cultural, morphological and biochemical characteristics. The aim of this study was to evaluate metabolic fingerprinting in the species-level identification of A. pleuropneumoniae isolates. Utilisation of carbon sources by these field isolates and six reference strains was characterised by the Biolog system (GN2 Microplate, MicroLog3 Version 4.20.05 software). Twenty-nine field strains were correctly identified by the Biolog system as A. pleuropneumoniae, 36 strains as A. lignieresii, two strains as H. paraphrohaemolyticus and one strain as A. equuli after 24 h of incubation. Among the six A. pleuropneumoniae reference strains the Biolog system identified one strain as A. pleuropneumoniae, four as A. lignieresii and one as H. paraphrohaemolyticus. There was no correlation between biotypes and serotypes of A. pleuropneumoniae and the carbon source utilisation pattern and species identification by the Biolog system. our data indicate that the efficacy of the Biolog system used here could be improved by including phenotypes of more A. pleuropneumoniae strains representing a wider geographical occurrence into the database.

Extraction and preliminary study of antibacterial compounds of three species of Aspergillus genus

In the interest of discovering new antibiotic molecules, the antibacterial activity of three fungal strains namely: Aspergillus quadrilineatus, Aspergillus niveus, and Aspergillus wentii isolated from particular ecosystems was sought against six bacterial strains including three with Gram-positive staining (Staphylococcus aureus, Bacillus subtilis, Enterococcus faecalis) and three with Gram-negative staining (Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae). The results of the agar cylinder technique highlighted that the three fungal strains showed a considerable antibacterial activity. In order to optimize the extraction conditions of the bioactive molecules, five solvents in different polarities were tested, of which chloroform turned out to be the best one. After the selection of this solvent, four culture media of different compositions were used in order to determine the most adequate medium for the production of antibacterial substances. The results revealed that Czapek-dox medium supplemented with yeast extract turned out to be the most favorable one for the production of bioactive molecules from both strains: A. quadrilineatus and A. niveus, while the most suitable medium for the A. wentii strain was Sabouraud. In addition, a study of the antibacterial effect of organic extracts by the Biolog micro-culture system was performed using a range of concentrations. The obtained results revealed that the extracts of the three fungal strains presented a remarkable activity with different concentrations and this, against all the tested bacterial strains. It was recorded only for the three used fungal species, the antibacterial activity was studied for the first time by the Biolog system.